Journal: Advanced Science

Article Title: Aged ZnO Nanoparticles Promote Breast Cancer Lung Metastasis by Remodeling the Pulmonary Microenvironment Through the CXCL10/Wnt/β‐Catenin Axis

doi: 10.1002/advs.78249

Figure Lengend Snippet: CXCL10 is upregulated in both macrophages and fibroblasts responding to aged ZnO NPs exposure. (A) Heatmaps exhibited multiplex cytokine profiling of serum and BALF from IT (PBS/fresh NPs/aged NPs intratracheal instillation) and IT + IV (IT exposure followed by intravenous injection of 4T1 cells) models ( n = 3). (B–E) Concentrations of significantly dyregulated cytokines in serum and BALF from mice exposed to ZnO NPs (fresh or aged), with or without subsequent 4T1 cell injection ( n = 3). (F) Venn plot showed the overlapping cytokines in serum and BALF between IT and IT + IV models. (G) ELISA detection of CXCL10 concentrations in serum and BALF of IT and IT + IV groups ( n = 3). (H) qPCR analysis of CXCL10 mRNA in lung tissue, primary fibroblasts, and alveolar macrophages post‐IT, normalized to GAPDH ( n = 3). (I, J) UMAP visualization (I) and dot plot (J) displaying CXCL10 expression across annotated cell types. (K, L) UMAP visualization (K) and dot plot (L) displaying CXCL10 expression across myeloid cell types. (M) Representative images of invasion and migration assays of 4T1 cells stimulated with 50 or 100 ng/mL murine recombinant CXCL10 (rCXCL10) for 72 h ( n = 3). (N) Quantification of invasion and migration assays ( n = 3). All data were represented as the mean ± SD. P values were calculated by one‐way ANOVA followed by Tukey's test. * p < 0.05, ** p < 0.01 vs control group; # p < 0.05, ## p < 0.01 vs fresh NPs group.

Article Snippet: CXCL10 (IP‐10) levels were measured using a mouse ELISA kit (Elabscience, E‐EL‐M0021) following the manufacturer's instructions.

Techniques: Multiplex Assay, Profiling, Injection, Enzyme-linked Immunosorbent Assay, Analysis, Tissue, Expressing, Migration, Recombinant, Control

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: Targeting Capability of D-EVs to Senescent NPCs is Mediated by the CXCL10-CXCR3 Axis. (A) Schematic diagram of RNA sequencing for senescent NPCs treated with D-EVs or not. (B-C) Volcano plots and heatmaps displaying differential gene expression in senescent NPCs treated with D-EVs or not. (D) A Venn diagram illustrates overlap between DEGs identified in senescent NPCs treated with D-EVs and key gene databases for cytokine-cytokine receptor interactions. (E) A Venn diagram illustrates overlap between DEGs identified in D-MSCs and chemokine signaling pathway key gene databases. (F-G) Network analysis of the hub genes among the above total DEGs. A protein-protein interaction network was created in the STRING database, while the Cytoscape software was used to determine the hub genes in the network. (H) GO analysis confirming enrichment of terms related to senescent NPCs treated with D-EVs or not in the BP categories. (I) GSEA analysis of cytokine-cytokine receptor interactions in D-EVs group versus TBHP group. (J) Western blot analysis confirmed the expression of CXCR3 in Control and senescent NPCs, and (S) quantitative analysis. (K) Western blot analysis confirmed the expression of CXCL10 in N-EVs and D-EVs, and (R) quantitative analysis. (L) Protein binding analysis of CXCR3 and CXCL10 by PyMOL. (M) Confocal analysis of the uptake of pHrodo-labeled EVs in senescent NPCs following the treatments of N-EVs, D-EVs with CXCL10 knockout/anti-CXCR3, or not, and (Q) quantitative analysis. (N) In vivo tracking of PKH26-labeled CXCL10 knockout D-EVs. (O) Flow cytometry showing uptake of different EVs with CXCL10 knockout/anti-CXCR3 or not by senescent NPCs, and (P) quantitative analysis. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against GAPDH (1:5000, 104941-AP, Proteintech), TSG101 (1:1000, DF8427, Affinity), CD9 (1:1000, AF5139, Affinity), CD63 (1:2000, 25682-1-AP, Proteintech), Calnexin (1:5000, 10427-2-AP, Proteintech), GM130 (1:20000, 11308-1-AP, Proteintech), CXCR3 (1:5000, 26756-1-AP, Proteintech), CXCL10 (1:2000, 10937-1-AP, Proteintech), MMP3 (1:2000, 17873-1-AP, Proteintech), ADAMTS5 (DF13268, Affinity), P16 (AF5484, Affinity), P21 (10355-1-AP, Proteintech), GPX4 (1:1000, 381958, Zen-bio), SLC7A11 (1:1000, 26864-1-AP, Proteintech), ACSL4 (1:5000, 22401-1-AP, Proteintech) and Tubulin (1:10000, T40103 , Abmart) overnight at 4 °C.

Techniques: RNA Sequencing, Gene Expression, Software, Western Blot, Expressing, Control, Protein Binding, Labeling, Knock-Out, In Vivo, Flow Cytometry

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: D-EVs Deliver GPX4 to Inhibit Ferroptosis in Senescent NPCs. (A) Representative Senescent-Tracker images of NPCs treated with N-EVs, D-EVs, Era, and D-Evs sh-CXCL10 . (B) Volcano plot of transcriptomic data comparing D-MSC and N-MSC. (C) KEGG pathway analysis of DEGs in D-MSCs versus N-MSCs. (D) Volcano plot of proteomic data comparing D-EVs and N-EVs. (E) KEGG pathway analysis of transcriptomic and proteomic data integration. (F) A Venn diagram illustrating the intersection of genes from the D-MSC transcriptome, the D-EVs proteome, and the ferroptosis-related gene set. (G) Bar graph showing the relative expression levels of core overlapping genes identified in (F). (H) MS analysis revealed that GPX4 is enriched in the D-EVs proteome. (I) Western blot analysis confirming GPX4 protein in D-EVs and N-EVs. (J) Western blot analysis of key senescence (p21, P16) markers in NPCs following treatment with PBS or N-EVs with CXCL10 or GPX4 knockout. (K) Representative images of EdU depicting cell proliferation ability in the control, TBHP, D-EVs, D-EVs sh-CXCL10 , D-EVs sh-GPX4 , and D-EVs sh-CXCL10+GPX4 groups. (L-M) Confocal images showing GPX4 delivery from different EVs to senescent NPCs at 12h and 24h co-culture, and (N) colocalization analysis. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against GAPDH (1:5000, 104941-AP, Proteintech), TSG101 (1:1000, DF8427, Affinity), CD9 (1:1000, AF5139, Affinity), CD63 (1:2000, 25682-1-AP, Proteintech), Calnexin (1:5000, 10427-2-AP, Proteintech), GM130 (1:20000, 11308-1-AP, Proteintech), CXCR3 (1:5000, 26756-1-AP, Proteintech), CXCL10 (1:2000, 10937-1-AP, Proteintech), MMP3 (1:2000, 17873-1-AP, Proteintech), ADAMTS5 (DF13268, Affinity), P16 (AF5484, Affinity), P21 (10355-1-AP, Proteintech), GPX4 (1:1000, 381958, Zen-bio), SLC7A11 (1:1000, 26864-1-AP, Proteintech), ACSL4 (1:5000, 22401-1-AP, Proteintech) and Tubulin (1:10000, T40103 , Abmart) overnight at 4 °C.

Techniques: Expressing, Western Blot, Knock-Out, Control, Co-Culture Assay